The skin has a protection function against the surrounding environment such as change of temperature or humidity, ultraviolet, and/or pollution materials. The normal function of the skin may be depressed by the excessive physical or chemical stimulation and stresses, or nutrient deficiency, causing skin-aging and/or skin-damage.
The external preparations for skin, such as cosmetics or topical formulations, are designed to maintain the skin in a beautiful and healthy state. There are a variety of cosmetics and/or topical formulations in the market, most of them are in the form of lotions, creams, detergents, dispersion solutions, or ointments. These skin preparations generally contain ingredients which may or may not have any dermatological effect on the skin. For example, to maintain the skin in a smooth, moisturizing condition, humectants, such as glycerin, sorbitol, propylene glycol, polysaccharide, and the like, are generally used in the cosmetics or topical formulations. None of these ingredients have any dermatological effect.
Also, for lightening or whitening the spots and freckles on the skin, which are generally formed due to abnormal melanine deposits on the skin, a regional application of ointment, cream or lotion containing kojic acid, vitamin C/ascorbic acid, glutathione, or cysteine can be useful. The use of hydroquinone and derivatives thereof, which are known as melanine generation inhibitors, are also commonly known for the purpose of bleaching erythema dyschromium perstans. Most of these ingredients, however, exhibit dermatological effect on inhibiting melanine formation.
In addition, vitamin A, soybean extract, and/or seaweed extract, which have been used in the external preparation for skin to provide an anti-aging effect, to improve wrinkle, and restore skin damage due to ultraviolet rays exposure, have exhibited both cosmetic and dermatological effect on skin.
However, the shortcomings of these compounds are that most of them are synthetic in nature and serve only narrow function, which not only may cause allergic reaction on skin but also may not be effective for skin conditions such as inflammation, wounds, and/or skin irritation. Thus, finding an ingredient that does not cause allergic reaction on skin but has multiple functions will be of great value to the dermatological and beauty field.
Ganoderma (Ganoderma lucidum Leyss ex Fr. Karst) is a polyporous fungus. It belongs to the class of Basidiomycetes, the family of Polypolaceae, and the genus of Ganoderma. In Chinese folklore, Ganoderma has been regarded as a panacea, which is probably due to certain efficacy of Ganoderma in treating many diseases. Some of the known medicinal or therapeutic effects of Ganoderma include treating patients with chronic bronchitis, chronic viral hepatitis, coronary heart disease, granulocytopenia, chronic Keshan disease, neurasthenia, progressive muscular dystrophy, atrophic myotonia and certain neurological diseases (See e.g., Liu et al., Chinese Medical Journal, 92:496–500 (1979)). There are also reports on Ganoderma as anti-HIV agent (See e.g., El-Mekkawy et al., Phytochemistry, 49: 1651–1657 (1998); Min et al., Chem. Pharm. Bull, 46: 1607–1612 (1998)), or for having anti-tumor, cardiovascular, antiviral, antibacterial, antiparasitic, and immune modulating activities (See e.g., Wasser et al., Critical Review in Immunology, 19:65–96 (1999)).
There are two major types of compounds found in Ganoderma which have been shown to be associated with the medicinal or therapeutic effects of Ganoderma. They are the polysaccharide compounds and the terpenoids. The polysaccharide compounds are primarily water-soluble. The terpenoids are oleaginous substances and are generally insoluble in water.
The polysaccharide compounds isolated from Ganoderma include heteroβ-glucans and their protein complexes (such as xyloglucans and acidic β-glucan-containing uronic acid, dietary fibers, lectins). The polysaccharides found in Ganoderma have been reported to possess anti-tumor and immune modulating effects (See Wasser et al., supra).
The Ganoderma terpenoids contain a lanostane skeleton. They are classified into several groups based on their carbon numbers and state of oxidation (Komoda et al., Chem. Pharm. Bull., 33:4829–4835 (1985)). These Ganoderma terpenoids include lanostanine-type triterpenoids (e.g., ganoderic acids A, B, C1, C2, D1, D2, E1, E2, F, G, H, I, J, K1, K2, L, Ma, Mb, Mc, Md, Me, Mf, Mg, Mi, Mj, Mk, Mn, N, O, P, Q, S, T, U, V, W, X, Y, and Z), 7-O-methyl-ganoderic acid O, trideacetyl ganoderic acid T, ganoderenic acids A, B, C, D, E, F, G, H, I, ganolucidic acids A, B, C, D, and E, lucidenic acids A, B, C, D1, D2, E1, E2, F, G, H, I, J, K, L, M, ganoderiol type 1 (A, B, F) and type 2 (C, D, E, F, G, H, and I), ganoderal A and B, epoxyganoderiol A, B, C, lucidone A, B, C, furanoganoderic acid, and other terpenoid components. Ganoderma terpenoids (e.g., ganoderic acids R, T, U-Z) have been reported to inhibit growth of hepatoma cells in vitro (See Toth et al., Tetrahedron Lett., 24:1081–1084 (1983)). The Ganoderma terpenoids are hereinafter referred to as “Ganoderma spore lipids” to reflect the water-insoluble, oleaginous characteristics of the compounds.
There have been reports on methods for breaking the epispores of Ganoderma spores. For example, Japanese Patent No. JP52041208 discloses an extraction method for breaking Ganoderma spores using mechanical force. Chinese Patent No. CN1134306 teaches a method for breaking the sporoderm of the Ganoderma spores by soaking the spores in water, followed by microwave-heating. Chinese Patent No. CN1165032 teaches a method for breaking the cell wall of Ganoderma lucidum spores by digesting the spores with skin-dissolving enzymes such as lysozyme, snail enzyme, cellulase, or hemicellulase, followed by ultrasonic breakage of the cell walls at 20–50° C. More recently, the inventor of this invention has disclosed a germination-activation method for treating the Ganoderma spores prior to the breaking of the epispores. See U.S. Pat. Nos. 6,316,002 and 6,468,542, which are herein incorporated by reference. The germination-activation procedure synchronizes the growth of and induces the maximum production of biological substances within of the Ganoderma spores. The germination-activated sporoderm-broken Ganoderma spores have demonstrated therapeutic effects on various diseases.
The biological substances in the Ganoderma spores contain, inter alia, active genes and promoters, active enzymes, sterols, cytokines, interferons, lactone A, ganoderma acid A, triterpenes, polysaccharides, vitamins, superoxide dismutases (SOD), glycoproteins, etc. These biological substances demonstrated superb medicinal effects, particularly on stimulating and modulating the nervous system and the immune system. These biological substances also demonstrated therapeutic effects on liver cancer and HBV infection. Additionally, when the Ganoderma spores were given to animals, the sporoderm-unbroken spores had an anti-tumor rate of 23.2%, which was substantially lower than the sporoderm-broken spores, which had an anti-tumor rate of 86.1%.
There have been reports on isolation or separation of the oleaginous substances (e.g., the terpenoids) from Ganoderma, most involving the use of organic solvents. For example, Min et al., Chem. Pharm. Bull., supra, disclose the isolation of lanostane-type triterpenes using column chromatography of a CHCl3-soluble fraction of the methanol extract of the Ganoderma spores. Lin et al., J. Chromatography, 410: 195–200 (1987) disclose the separation of oxygenated triterpenoids from Ganoderma lucidum by high-performance liquid chromatography of a methanolic extract of Ganoderma lucidum. These methods are unsatisfactory due to complex extraction procedures and low yield of the oleaginous substances.
Recently, a method for extracting oleaginous substances from sporoderm-broken Ganoderma spores has been disclosed also by the inventor of this invention in U.S. Pat. No. 6,440,420, which is herein incorporated by reference. The method includes (1) breaking up the sporoderm of the germination-activated Ganoderma spores by a mechanical means; and (2) extracting the Ganoderma spore lipids from the sporoderm-broken germination-activated Ganoderma spores by a supercritical fluid carbon dioxide (SCF-CO2) extraction method. This method produces high yield of Ganoderma spore lipids from Ganoderma (i.e., the yield of the Ganoderma spore lipids is about 37% by weight of the entire biological substances released from Ganoderma). The Ganoderma spore lipids extracted by this method is transparent and odorless.
In the present invention to be described in the following sections, an external preparation for skin comprising the Ganoderma spore lipids extracted from sporoderm-broken Ganoderma spores using the SCF-CO2 method is provided. The Ganoderma spore lipids demonstrate multiple functions in smoothening skin, reducing wrinkles, defying aging, lessening inflammation, whitening pigmentation, and alleviating skin irritation, thus making it an ideal ingredient to be used in both the cosmetic and the dermatological field.